A 24-well Transwell chamber (pore size, 8 mm) with or without Matrigel coating (BD Biosciences) was used to detect tumor cell invasion and migration, respectively. Tumor cells (1×105) co-cultured with or without TAMs were plated in the upper chamber, and different conditional media (CM) containing 10% FBS (Thermo Fisher Scientific, Inc.) with or without U0126 [1,4-diamino-2, 3-dicyano-1,4-bis (2-aminophenylmercapto) butadiene; ERK1/2 inhibitor; 10 µg/ml; Cell Signaling Technology, Inc.] were added to the lower chamber. Once the cells were incubated for 24 h, the non-penetrated cells were scraped from the upper surface of the filter, and the penetrated cells, representing invaded or migrated cells, were stained with crystal violet solution (Sigma-Aldrich; Merck KGaA) at room temperature for 10 min. Cells were counted under a phase microscope, photographed and analyzed using ImageJ 1.42 software (National Institutes of Health, Bethesda, MD, USA).
Erk1 2 inhibitor
Manufactured by Cell Signaling Technology
The ERK1/2 inhibitor is a laboratory reagent used to study the extracellular signal-regulated kinase (ERK) signaling pathway. It functions by inhibiting the enzymatic activity of the ERK1 and ERK2 proteins, which are key components of the mitogen-activated protein kinase (MAPK) cascade.
Automatically generated - may contain errors
Lab products found in correlation
Matrigel coating, by BD (1 mentions)
Transwell chamber, by BD (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Crystal violet solution, by Merck Group (1 mentions)
Nf κb inhibitor bay 11 7082, by Merck Group (1 mentions)
Ly204002, by Merck Group (1 mentions)
Ns 398, by Cayman Chemical (1 mentions)
Wortmannin wm, by Merck Group (1 mentions)
Pge1 oh, by Cayman Chemical (1 mentions)
U0126, by Cell Signaling Technology (1 mentions)
2 protocols using erk1 2 inhibitor
1
Transwell Assay for Tumor Cell Invasion and Migration
2
Pharmacological Modulation of COX-2 and EP4 Signaling
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NS398 (COX-2 inhibitor) was purchased from Cayman Chemical (Michigan, MI). ONO-AE3-208 (selective EP4 antagonist) was a gift of ONO Pharmaceuticals, Osaka, Japan. PGE2 (EP1-4 ligand) and PGE1OH (EP4 agonist) were purchased from Cayman Chemical. Wortmannin (WM), an irreversible PI3K inhibitor, and LY-204002 (LY), a reversible PI3K inhibitor all purchased from Sigma-Aldrich, ERK1/2 inhibitor U0126 from Cell Signalling, MA. NF-κB inhibitor, Bay 11-7082 (Sigma-Aldrich, ON, Cat # B5556) is a kind gift of Dr. Xiufen Zheng (Department of Pathology and Laboratory Medicine, University of Western Ontario). For all treatments in vitro or in vivo, respective vehicles served as the control.
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